GSM7118540: sample1_control1 (biol rep 1); Homo sapiens; RNA-Seq
Freshly isolated primary human classical monocytes (CD14+CD16-) from two donors were cultured for 8 hours in triplicates in cell repellent plates in the presence of control or Wnt-3a conditioned media.   Media were comprised of 80% L- or L-Wnt-3a- CM and 20% RPMI-1640 (both containing 10% FBS and 1% Penicillin/Streptomycin) at 37°C in 5% CO2. L cells and L-Wnt-3a cells were propagated according to the ATCC guidelines and CM were produced according to the ATCC protocols. Total RNA was extracted using the Direct-zol RNA MiniPrep kit. Replicates of high RNA integrity (RIN ≥ 7.8) were processed. Libraries were prepared using the NEBNext Ultra II RNA Library Prep kit with the NEBNext Poly(A) mRNA Magnetic Isolation Module, starting with ~ 500 ng of total RNA. Ten PCR cycles were performed during library amplification. Libraries were quantified by Qubit and TapeStation and sequenced on a NextSeq 500 instrument using a NextSeq 500/550 High Output Kit v2.5 (75 Cycles) kit.